EAGER: Toward determining the key interactions between the SARS-CoV2 virus components and host cells

该项目将带来严格的生物物理知识，以选择对人类健康，活动和经济福祉具有灾难性影响的SARS-CoV 2病毒的蛋白质组分。 病毒感染周期中的关键事件包括病毒与易感宿主细胞的相互作用以及感染后的各种细胞反应。将进行实验研究，以表征参与这些阶段的蛋白质。通过结合两个研究小组广泛的热力学和结构专业知识，将描述和量化关键的结构特征和分子相互作用。这些数据将提供一个框架，以测试基于细胞的感染性试验中的发现。 两名研究生将接受培训，该项目的结果将用作针对生物物理本科生和研究生的冠状病毒研讨会课程的基础，以培养未来在该领域具有经验的劳动力。SARS-CoV-2与宿主之间的关键相互作用是病毒刺突跨膜蛋白的受体结合结构域（RBD）与人ACE 2跨膜蛋白的S1结构域之间的相互作用。结合是感染所必需的，决定了特定病毒可以感染的宿主范围。该项目将定量这种关键的相互作用，确定病毒-宿主特异性概况，测试导致SARS-CoV-2大流行的起源和人畜共患病事件的各种机制，并产生具有良好稳定性和特异性的可溶性模拟物，可用于限制或预防感染。此外，三个辅助蛋白，Orf 3a，Orf 7a和Orf 8，干扰或劫持宿主细胞过程的特点，结构和功能，使用各种光谱和流体动力学方法。为了进一步探索冠状病毒的进化起源及其宿主特异性，祖先序列重建方法和共识设计将应用于这些靶点。EAGER奖由分子和细胞生物科学部的分子生物物理学计划颁发，使用冠状病毒援助，救济，该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This project will bring rigorous biophysical knowledge to select protein components of the SARS-CoV2 virus that has catastrophic effects on human health, activity, and economic well-being. Critical events in the viral infection cycle include interaction of the virus with susceptible host cells and a variety of cellular responses post-infection. Experimental studies will be performed to characterize proteins that participate in these stages. By combining the extensive thermodynamic and structural expertise of two research groups, key structural features and molecular interactions will be described and quantified. The data will provide a framework to test the findings in cell-based infectivity assays. Two graduate students will be trained, and the results of this project will be used as the basis for a seminar course on coronaviruses directed at biophysics undergraduate and graduate students to develop future workforce with experience in this area. A critical interaction between SARS-CoV-2 and host is that between the receptor binding domain (RBD) of the viral spike transmembrane protein and the S1 domain of the human ACE2 transmembrane protein. Binding is required for infection, determining the host range that a particular virus can infect. This project will quantify this critical interaction thermodynamically, determine the virus-host specificity profile, test various mechanisms for the origin and zoonotic events that led to the SARS-CoV-2 pandemic, and generate soluble mimics with favorable stabilities and specificities that can be used to limit or prevent infection. In addition, three accessory proteins, Orf3a, Orf7a, and Orf8, that interfere with or hijack host cellular processes will be characterized, both structurally and functionally using various spectroscopic and hydrodynamic methods. To further probe the evolutionary origins of coronaviruses and their host specificities, ancestral sequence reconstruction methods and consensus design will be applied to these targets.This EAGER award is made by the Molecular Biophysics Program in the Division of Molecular and Cellular Biosciences, using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.